Ultra-performance Liquid Chromatography Method Research Articles

Profiling the occurrence of biogenic amines in wine from Chinese market and during fermentation using an improved chromatography method

The determination of biogenic amines (BAs) plays an important role in ensuring food safety. In this study, the occurrence of BAs in 98 wine samples readily available to Chinese consumers, as well as the dynamics of BAs during the spontaneous fermentation of Cabernet Sauvignon and Marselan wines were determined by an improved ultra-performance liquid chromatography (UPLC) method. It was found that this novel approach presented a reduced Dns-Cl amount (0.55 mL), lower limits of detection (1.5–4.2 μg/L) and quantification (4.9–14.1 μg/L), and a much shorter analysis time (6 min). The cumulative BA levels varied significantly, from 3.46 ± 0.05 mg/L in Sauvignon Blanc to 59.12 ± 0.09 mg/L in Cabernet Sauvignon. Putrescine was the most abundant amine in wine samples, with concentrations ranging from 0.32 ± 0.12 mg/L in Cabernet Sauvignon to 31.17 ± 0.27 mg/L in Pinot Noir, while spermine was not detected in 80% of the wine samples and had the lowest concentration (average 0.205 ± 0.13 mg/L). None of the investigated samples presented relevant levels of histamine and tyramine above the limits specified by the European Food Safety Authority (EFSA). Thus, wines available in China are not a serious toxicological risk regarding the type and content of BAs. The malolactic fermentation (MLF) and the later stage of alcohol fermentation (AF) were found to be critical control points (CCPs) in the spontaneous fermentation of both Marselan and Cabernet Sauvignon wines. Moreover, the contents of putrescine, tyramine and tryptamine were significantly correlated with the contents of other amines, and, thus, could be used as important indicators of the quality and safety of wines. In general, these results are helpful to assess the risk of dietary intake of BAs and to evaluate the winemaking processes by food safety authorities. ● A rapid and high resolution UPLC method was optimized to determine biogenic amines. ● Abundant putrescine content was found in wine samples. ● Wines available in China may be considered safe for consumption. ● MLF and later stage of AF were the critical control points of red wine fermentation. ● Putrescine, tyramine and tryptamine were used as important indexes of wine safety.

Polyphenol Enriched Chrysanthemum morifolium Extract Increase Energy Expenditure and Decrease Hepatic Lipid Accumulation in Rats Fed a High Fat Diet

ObjectivesThis study aimed to examine the improvement effects of polyphenol enriched Chrysanthemum morifolium extract (CE) on energy expenditure and hepatic lipid accumulation in rats fed a high-fat diet. MethodsThe polyphenols identified in CE were analyzed by ultra-performance liquid chromatography (UPLC) method. Male Sprague-Dawley rats were randomly divided into four groups. Each group fed a normal diet (NOR), 45% high-fat diet (HF), or HF supplemented with 0.2% CE (CEL), or HF supplemented with 0.4% CE (CEH) for 13 weeks. ResultsPolyphenolic compounds of CE were confirmed that three flavonoids, such as luteolin-7-O-glucoside, luteolin-7-O-glucuronide and apigenin-7-O-glucoside, and three phenolic acids, including chlorogenic acid, 1,5-Dicaffeoylquinic acid and 3,5-Dicaffeoylquinic acid. CE significantly increased oxygen consumption (VO2), carbon dioxide production (VCO2) and energy expenditure (EE) compared with HF. CE supplementation was lower the serum concentrations of total cholesterol (TC), triglyceride (TG), non-esterified fatty acids (NEFAs) and low-density lipoprotein cholesterol (LDL-C), and decreased the levels of hepatic total lipids, TC and TG. Hepatic lipogenic gene expression such as cluster of differentiation 36 (CD36), sterol regulatory element binding protein-1c (SREBP-1c), ATP citrate lyase (ACLY), acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), stearoyl-CoA desaturase 1 (SCD1), and diacylglycerol O-acyltransferase 2 (DGAT2) significantly down-regulated by CE supplementation. The mRNA expression involved in fatty acid oxidation such as peroxisome proliferator-activated receptor alpha (PPARα) and carnitine/palmitoyl-transferase 1α (CPT1α) were up-regulated in the CEH group than HF group. Hepatic adenosine monophosphate-activated protein kinase (AMPK) activity was significantly increased in the CE groups than HF group. ConclusionsIt is suggested that the polyphenol enriched CE might increase energy expenditure and reduce hepatic fat accumulation via modulating hepatic lipogenic and fatty acid oxidation gene expression with increasing AMPK activation. Funding SourcesThis research was supported by the BK21 FOUR (Fostering Outstanding Universities for Research) funded by the Ministry of Education (MOE, Korea) and National Research Foundation of Korea (NRF).

Comparison of the Purity and Impurity of Glucagon-for-Injection Products under Various Stability Conditions

Glucagon is a polypeptide hormone that serves as an essential therapeutic agent in the emergency treatment of hypoglycemia. Recently, the first generic glucagon for injection was approved. However, unlike its brand name counterpart, which is produced via recombinant DNA, the generic glucagon is produced using a chemical synthesis method. Regardless of its origin, impurities may occur in both glucagon drug products. While these impurities may greatly compromise the safety and efficacy of the glucagon drug products, studies accessing the impurities of glucagon for injection are limited. This manuscript analyzed the stability and impurities of a generic and brand glucagon for injection, including desamido and non-desamido impurities, under various storage and temperature conditions using an ultra-performance liquid chromatography method. The glucagon products were analyzed after 6 and 24 months of storage under room temperatures (20–25 °C). In addition, the products were also assessed after 6 months of storage under high temperatures (40 °C). Under each stability storage condition, three lots of the synthetic glucagon were evaluated by UPLC with at least one lot of the recombinant glucagon for comparison. A total of 37 peaks were identified (except for the solvent peaks, which appeared at retention times less than 1.5 min) from the synthetic and recombinant glucagon lots. It was found that the number of impurities observed in the synthetic glucagon were lower than the referenced recombinant glucagon across all stability conditions. Throughout all tested conditions, the synthetic glucagon for injection had an averaged purity of 92.8–99.3%, while the referenced recombinant drug had an averaged purity of 70.3–91.7%. Based on the study results, it can be concluded that the impurity profile for the synthetic glucagon for injection has a comparable and even lower level of impurities than the recombinant version under all stability conditions.

Forced Degradation Behaviour with a Developed and Validated RP-UPLC Method for Estimation Lamivudine and Dolutegravir in Combined Pharmaceutical Dosage Form

A simple, fast, precise, specific, and accurate reversed phase Ultra Performance Liquid Chromatographic (UPLC) method was developed and validated for the forced degradation studies of the Lamivudine and Dolutegravir in tablet dosage form. Chromatogram was run HSS C18 (2.6 x 50mm, 1.6µm). Mobile phases containing 70% 0.01N disodium hydrogen phosphate: 30% Methanol). The elution of analytes was achieved with a flow rate at 0.3 mL/min. Potassium dihydrogen phosphate was used as a buffer in this experiment. Temperature was maintained at 30°C. Optimized wavelength selected at 260nm. The detector response was linear in the concentration range of 25-150μg/mL respectively. Retention time of Lamivudine and Dolutegravir were found to be 1.408min and 1.739min. %RSD of the Lamivudine and Dolutegravir were and found to be 0.8 and 0.8. The % recovery was obtained as 100.39% and 100.37% for Lamivudine and Dolutegravir. LOD, LOQ values obtained from regression equations of Lamivudine and Dolutegravir were 0.41, 1.25 and 0.09, 0.26. Regression equation of Lamivudine was y = 24270x + 12218 and for Dolutegravir was y = 34783x + 1060. Since retention times and run times were reduced, the method established was easy and premium and it could be used in periodic quality control tests in industries.

Method development for PPB culture screening, pigment analysis with UPLC-UV-HRMS vs. spectrophotometric methods, and spectral decomposition-based analysis

PPB carotenoids are usually measured through spectrophotometric analysis, measuring total carotenoids (TCs) which has low accuracy and cannot identify individual carotenoids or isomers. Here, we developed an ultra-performance liquid chromatography method with ultraviolet and high-resolution mass spectrometry detection (UPLC-UV-HRMS) to quantify neurosporene, lycopene, and bacteriochlorophyll a contents in PPB cultures. The method exhibited satisfactory recoveries for individual pigments (between 82.1% and 99.5%) and was applied to a range of mixed PPB cultures. The use of a C30 column also enabled the detection of three different isomers of lycopene. In addition, a method for anaerobic photoheterotrophic PPB cultivation to acquire live-cell spectrophotometric information was developed and tested by modifying a standard microbial culture microplate system. A rapid, and relatively low effort principal component analysis (PCA) based decomposition of the whole-cell spectra for pigment analysis in the microplates was also developed. Analysing whole-cell spectra via PCA allowed more accurate prediction of individual pigments compared to absorption methods, and can be done non-destructively, during live-cell growth, but requires calibration for new media and microbial matrices.

A Validated Ultra Performance Liquid Chromatography Method for Simultaneous Estimation of Diacerein and Aceclofenac in Bulk and Pharmaceutical Formulation

A simple, exceptionally cost-effective, extremely accurate, quite precise and highly reproducible analytical method was developed and validated for simultaneous estimation of Diacerein and Aceclofenac in bulk and Pharmaceutical formulation. The new Ultra Performance Liquid Chromatography technique was developed. Method- The separation has been done on column ACQUITY UPLC BEH Shield C18 (50 x 2.1mm), 1.7µm (40°C temperature). The mobile phase contained Buffer and Acetonitrile (Buffer: ACN) (55:45 V/V).. The flow rate was set at 0.4ml/min, and detected at 268nm with PDA detector. Mobile phase was sonicated for 15 min. before use. 5μl of samples of standard stock solution and tablet solution were injected. Different trials were performed to separate diacerein and aceclofenac. The total run time of the detection was 4 min. The developed method was validated against different validation parameters. Results - The retention times were obtained at 1.762min. and 2.891 min. for Diacerein and Aceclofenac respectively. Accuracy study was observed in the range of 100.33% to 101.67% with less than 2% RSD. LOQ was found to be 2.98μg/ml and 5.9041μg/ml. similarly LOD was found to be 0.9841μg/ml and 1.9467μg/ml. for Diacerein and Aceclofenac respectively. Precision %RSD for intraday and interday were found to be 0.19 and 0.18 for Diacerein , 0.44 and 0.16 for Aceclofenac respectively. Linearity was found in the range of 2.5-17.5μg/ml and 5-35μg/ml for Diacerein and Aceclofenac respectively. The method was found robust by deliberate changes in the flow rate, ratio of mobile phase and detection wavelength. Conclusion-The method was found to be satisfactory and can be used successfully for determination of Diacerein and Aceclofenac simultaneously in bulk and pharmaceutical dosage form.

Development of a New Validated Stability Indicating Method for Quantification of Fenofibric acid and Pitavastatin by Ultra Performance Liquid Chromatography

The main objective of the present work is to develop an efficient, unique, reliable Ultra performance liquid chromatographic method for the simultaneous quantificaiton of Fenofibric acid and Pitavastatin in bulk and Pharmaceutical formulations. The Chromatographic separation of the selected combination of drugs was performed on a Kinetex C8 column (150mm x4.6mm, 2.6 µ) using an isocratic elution with a buffer containing 0.1% formic acid and acetonitrile at a ratio of 80:20 as a mobile phase with a flow rate of 0.5 mL/min at ambient temperature and wavelength at 266nm.The method produced reliable results at optimised chromatographic conditions. Analysis of honest linearity within a concentration range of 135- 2025 µg/mL of fenofibric acid and 2-30 µg/mL of Pitavastatin with regression coefficients of 0.9992 and 0.9997 respectively was obtained. The retention time for fenofibric acid and pitavastatin were 1.778 min and 3.171 min respectively. LOD and LOQ were found to be 1.35 µg/mL, 0.02 µg/mL and 13.5 µg/mL, 0.2 µg/mL for fenofibric acid and pitavastatin respectively.The proposed method was validated in accordance with the guidelines of the International Council for Harmonization (ICH). All the obtained validation results of the proposed method were found to satisfactory and was succesfully applicable to the analysis of the bulk and the pharmaceutical formulations.

Nitrites as precursors of N-nitrosation in pharmaceutical samples – A trace level analysis

The ultra-performance liquid chromatography (UPLC) method, which involves pre-column derivatization of nitrite with 2,3-diaminonaphthalene (DAN) to form 2,3-naphthotriazole (NAT), offers the advantages of easy sample preparation, simple derivatization, stable derivatives, rapid analysis, high sensitivity and specificity and lack of interferences for determining nitrite in pharmaceutical samples. Determination of NAT was performed on a an Acquity UPLC HSS T3 column using a gradient elution of 0.1% formic acid with acetonitrile at flow rate of 0.4 mL/min and temperature at 45 °C. The single-quadrupole mass detector was operated in the positive ion mode. Quadrupole mass analyser was employed in selected ion monitoring mode using a target ion at m/z = 170 as [M+H]+. The UPLC-MS method was validated as per International Council on Harmonization (ICH) guidelines in terms of linearity, limit of detection, limit of quantification, selectivity, accuracy, precision, intermediate precision and stability. The UPLC-MS method was demonstrated to be applicable for the determination of nitrite in various pharmaceutical samples. The proposed UPLC-MS method was used to study the effect of nitrite content in pharmaceutical products on the formation of N-nitrosamines. The high importance of nitrites in relation to the N-nitrosation reaction was discussed. As deduced from theory and justified by the presented results, reducing the nitrite concentration could definitely solve the N-nitrosamine contamination. Nitrites, unlike secondary and tertiary amines, are universal precursors to any N-nitrosamine, so this solution is easily transferable to any relevant pharmaceutical product.

A Chemometric Strategy in the Development of a RP-UPLC Method for the Quantitative Resolution of a Two-Component Syrup Formulation.

A novel chemometric strategy was implemented in the development of a new ultraperformance liquid chromatography method for the quantitative estimation of guaifenesin and pseudoephedrine hydrochloride in a two-component syrup formulation with minimal experimental effort, time and reagent. A full factorial design with three factors was investigated to find optimal working conditions of chromatographic factors (column temperature, flow rate, and 0.1M H3PO4% in mobile phase) that affect the chromatographic separation. Then, optimum experimental conditions providing adequate separation of the analyzed drug substances within the short runtime were determined. Under optimal experimental conditions, the retention times for guaifenesin and pseudoephedrine hydrochloride were obtained as 0.817 and 1.430min, respectively. In the optimized RP-UPLC method, chromatographic response was reported as a linear function of concentration between 5.0 and 80.0μg/mL for guaifenesin and 10.0-90.0μg/mL for pseudoephedrine hydrochloride. The proposed method was carefully validated and successfully applied to quality control and analysis of a cough syrup preparation containing guaifenesin and pseudoephedrine hydrochloride. Consequently, the proposed reversed-phase ultraperformance liquid chromatography method provided an opportunity to quantify relevant drugs with small amount of reagents and short runtime.

Development and Validation of Ultra Performance Liquid Chromatography (UPLC) Method for the Quantitative Estimation of Caffeine in Non-Alcoholic Soft and Energy Drinks.

The popularity of beverages such as cold coffee, iced tea, and energy drinks has risen tremendously among athletes and youths. Energy drinks are used to enhance performance due to the presence of a high amount of caffeine (CFN) and sugars, as well as other constituents such as vitamins, amino acids, taurine, extracts of Ginkgo biloba, ginseng, guarana, and other herbal products. Commercial drinks are promoted as being beneficial to health; thus it is an important concern regarding adverse effects linked with these drinks or products. The aim of the study is to develop and validate the ultra performance liquid chromatography-photodiode array detector (UPLC-PDA) method for the estimation of CFN in eight marketed non-alcoholic drinks including two soft drinks and six energy drinks. The chromatographic separation of CFN was achieved on Acquity UPLC® CSHTM C18 1.7 µm, 2.1 × 100 mm column, using isocratic mode, mobile phase comprising acetonitrile and water (30:70, v/v) at a flow rate 0.3 mL/min using injection volume 1 µL. The elution of CFN occurred at 1.06 min, and the calibration curve of the CFN was computed from the peak area ratio detected at 273 nm. All the validation parameters were found within the assay variability limits as per ICH guidelines. The obtained results revealed that the soft drinks SD1 and SD2 have 87.21 ± 1.28 and 101.81 ± 1.52% (w/w), whereas CFN concentration in energy drink brands ED1, ED2, ED3, ED4, ED5, and ED6 was 95.90 ± 1.62, 64.0 ± 1.07, 76.68 ± 1.95, 74.97 ± 2.33, 82.09 ± 2.43, and 88.04 ± 2.94% (w/w) of labeled claims, respectively. The developed UPLC method was found suitable for the quality control of commercial soft and energy drinks containing CFN. The developed chromatographic method is very simple, cost effective and could be utilized for the routine analysis of caffeine in the soft and energy drinks.

Quantification of Lipid and Peptide Content in Antigenic Peptide-loaded Liposome Formulations by Reversed-phase UPLC using UV Absorbance and Evaporative Light Scattering Detection

Antigenic peptide-loaded cationic liposomes have shown promise as cancer vaccines. Quantification of both peptides and lipids is critical for quality control of such vaccines for clinical translation. In this work we describe a reversed phase ultra-performance liquid chromatography (RP-UPLC) method that separates lipids (DOTAP, DOPC and their degradation products) and two physicochemically different peptides within 12 min. Samples were prepared by dilution in a 1:1 (v/v) mixture of methanol and water. Peptide quantification was done via UV detection and lipids were quantified by an evaporative light scattering detector (ELSD), both coupled to the RP-UPLC system, with high precision (RSD < 3.5%). We showed that the presence of lipids and peptides did not mutually influence their quantification. Limit of detection (LOD) and limit of quantification (LOQ), as determined in the ICH guidelines, were 6 and 20 ng for DOTAP, 12 ng and 40 ng for DOPC, 3.0 ng and 8.0 ng for peptide A and 2.4 ng and 7.2 ng for the more hydrophobic peptide B. Finally, lipid degradation of DOTAP and DOPC was monitored in peptide loaded DOTAP:DOPC liposomes upon storage at 4 °C and 40 °C.

VETERİNER DOZAJ FORMUNDA İKİ ANTELMİNTİK İLACIN EŞZAMANLI KANTİTATİF ANALİZİ

In this research article, a new chromatographic method ultra-performance liquid chromatography (UPLC) method was developed for the quantification of ivermectin (IMT)-preziquantel (PRZQ) in a two-component mixture and veterinary dosage form mixture. The separation of the studied veterinary drugs (IMT and PRZQ) was carried out on Waters Acquity ® BEH C18 column (50mm x 2.1 mm i.d., 1.7 m) using a mobile phase consisting of water-acetonitrile-methanol (10:70:20, v/v). In the chromatographic analysis, flow rate was 0.38 mL/min, and column temperature was maintained at 42 0C. The chromatographic detection of IMT and PRZQ was made at the wavelength of 220.0 and 245.0 nm, respectively. The developed UPLC method was validated and applied to real samples consisting of IMT and PRZQ. Recovery assay results were found to be 100.4 % for PRZQ and 101.8 % for IMT. The presented UPLC method is introduced as an alternative choice for the quality control of pharmaceutical preparations containing these drugs. For the quality control of pharmaceutical preparations containing these drugs, the presented UPLC method is introduced as an alternative experimental analysis choice.

Stability-Indicating New RP-UPLC Method for Simultaneous Determination of a Quaternary Mixture of Paracetamol, Pseudoephedrine, Chlorpheniramine, and Sodium Benzoate in (Cold-Flu) Syrup Dosage Form.

A combination of paracetamol, pseudoephedrine, chlorpheniramine, and sodium benzoate in (Cold-Flu) 1,2,3 Syrup dosage form is specified for the treatment of common cold and flu symptoms. The functional role of this study is to develop a novel, reliable, and selective stability-indicating reversed-phase ultra-performance liquid chromatography (RP-UPLC) method for simultaneous identification of a quaternary mixture of paracetamol, pseudoephedrine, chlorpheniramine, and sodium benzoate in (Cold-Flu) 1,2,3 Syrup dosage form. The specific method is accomplished using an Acquity UPLC HSS T3 C18 column (2.1 mm × 100 mm), 1.8 µm particle size with pore size 100 Å, utilizing a mixture of purified water-methanol-trifluoroacetic acid (72.5:27.5:1.5, v/v) as the mobile phase at a flow rate of 0.3 mL/min. The column void volume is 1.15 min. UPLC detection is adjusted at 205 nm using a photodiode array detector. Calibration curves are obtained in the linearity ranges: 25-500 µg/mL for paracetamol, 10-50 µg/mL for pseudoephedrine, 0.5-5 µg/mL for chlorpheniramine, and 3-30 µg/mL for sodium benzoate with a correlation coefficient > 0.9992. The mean recovery of the developed method is tested and shows good recovery results between 99-101%; selectivity and forced degradation studies are investigated as per the International Council for Harmonisation Guidelines and no interference is detected due to degradation peaks. The proposed stability-indicating UPLC method for simultaneous determination of the three drugs, paracetamol, pseudoephedrine, and chlorpheniramine, with a preservative sodium benzoate in (Cold-Flu) 1,2,3 Syrup dosage form is successfully accomplished, developed, and validated, and can be easily used in the analysis of drugs in pure or dosage form. The novelty of the current research work lies in the development of the UPLC method for simultaneous determination of a quaternary mixture of paracetamol, pseudoephedrine, chlorpheniramine, and sodium benzoate in (Cold-Flu) 1,2,3 Syrup dosage form.

Development and Validation of Novel Rapid Stability Indicating Mass Compatible UPLC Method for the Simultaneous Estimation of Assay and Related Substances of Remdesivir in Bulk and Injectable Dosage Form

Developed and validated a simple, rapid, new mass compatible reverse phase ultra-performance liquid chromatography (UPLC) method for the simultaneous estimation of assay and related substance of remdesivir in bulk and injectable dosage form. The developed method is capable of separating all the potential process and degradation impurities within 15 min. Chromatography was carried out using Waters Acquity BEH C18 column (50 mm length, 2.1 mm internal diameter and 1.7 μ particle size) with gradient elution. Ghost buster column (30 mm length, 2.1 mm internal diameter) was used as inline filter to suppress the response of peaks originating from mobile phase. Mobile phase A composed of 10 mM ammonium acetate buffer pH 3.5 and acetonitrile in the ratio of 90:10 v/v and mobile phase B composed of 10 mM ammonium acetate buffer pH 3.5 and acetonitrile in the ratio of 10:90 v/v. The flow rate of the mobile phase was set at 0.35 mL/min and the column temperature was maintained at 35 ºC. The detection was carried out using UV detector at 245 and 270 nm. Method validation was performed according to ICH guidelines. The validated method is simple, rapid, mass compatible, economical, specific, accurate, linear, precise, rugged, sensitive, robust and stability indicating. This method can be employed for the simultaneous estimation of assay and related substance of remdesivir in bulk and injectable dosage forms at quality control laboratories of pharmaceutical industries.

Development of Ultra performance Liquid Chromatography (UPLC) method for the Separation of Alfuzosin Hydrochloride and its organic impurities

For assessing alfuzosin hydrochloride in the presence of organic contaminants and degradants by forced breakdown products, a novel, quick, precise, sensitive, specific, and stability-indicating reverse-phase (RP-UPLC) approach was established. Acquity BEH C18 (50 × 2.1 mm,1.7), BEH C8 (50 × 2.1 mm,1.7), Acquity CSH C18 (50 × 2.1 mm,1.8), and Acquity BEH C18 (100 × 2.1 mm,1.7) are among the rapid liquid chromatography columns that have been investigated. Low and neutral pH buffers were also used. Furthermore, the best chromatographic separation was achieved with a Waters Acquity HSS T3 C18, 100 mm × 2.1 mm, particle size 1.8, UV detection at As a mobile phase carrier, (acetonitrile, tetrahydrofuran) is used. The final technique conditions enable excellent impurity separation, which can be used to analyse the stability of alfuzosin hydrochloride commercial samples as well as for quality control throughout production. The gradient time was about 6.5 min, and the re-equilibration time was roughly 1.5 min, allowing for more than 100 samples each day to be analysed. Aside from that, the pH sensitive analytical approach addressed in both the US and European Pharmacopoeias was explored.

PENGARUH MINYAK KELAPA MURNI DAN MINYAK INTI SAWIT TERHADAP PERTUMBUHAN RAMBUT PADA KELINCI

Hair is something that comes out of the skin and has a shape like a thin thread. Hair has many things, including protection from the sun's function, adding beauty and color lines to the face, protecting the eyes from sweat, dirt and dust, and others. Hair is composed of a protein substance called keratin. To determine the VCO and PKO in the rate of growth activity and hair distribution rate. Extraction of PKO from palm kernel and VCO from the coconut fruit, characterization testing including fatty acids and squalene using the Gas Chromatography (GC) method, free fatty acids using the titration method with 0.01N KOH, and finally vitamin E using the Ultra Performance Liquid Chromatography (UPLC) method. smeared every 2 times a day. Tests for the distribution rate of hair growth and daily hair growth rate were carried out every 5th, 10th, 15th, 20th, 25th, 30th day. And weight considerations were made on the 30th day. compared to Pure Coconut Oil in hair growth in rabbits.

Multilayered solid phase extraction and ultra performance liquid chromatographic method for suspect screening of halogenated pharmaceuticals and photo-transformation products in freshwater - comparison between data-dependent and data-independent acquisition mass spectrometry

Since conventional biological wastewater treatments are not admittedly effective to convert pharmaceutical active compounds (PhACs) into nontoxic products, natural abiotic mechanisms such as solar photolysis arises as an important degradation process, especially for halogenated molecules. In the present work, photolysis simulation was carried out in-lab for precursors and their respective photo-transformation products (photo-TPs), which were analyzed through reversed-phase ultra-high performance liquid chromatography coupled to high resolution mass spectrometry (RP-UHPLCHRMS). An in-house library was created in order to provide reference information for target (precursors) and suspect screening (photo-TPs) analysis of freshwater samples from impacted aquatic environments. Strategies in the use of data-dependent acquisition (DDA) and data-independent acquisition (DIA), as well as the data processing software are discussed here for the identification of 6 PhACs and photo-TPs. Because no standards of photo-TPs were available, only the target compounds, i.e. sitagliptin (398 ± 2 ng L−1), iohexol (209 ± 5 ng L−1), lamotrigine (103 ± 10 ng L−1), losartan (43 ± 10 ng L−1), ofloxacin (28 ± 7 ng L−1), and sertraline (25 ± 7 ng L−1) could be quantified through multiple standard additions.

Partial Validation of Ultra Performance Liquid Chromatography Method for Quantification of Isoniazid-Pyrazinamide in Human Samples

Isoniazid (INH) and pyrazinamide (PZA) are first-line drugs in the treatment of meningitis of tuberculosis, in addition to rifampicin. The use of these drugs will determine the success of therapy to kill Mycobacterium tuberculosis in brain tissue that is difficult to penetrate by other drugs. Therefore, it is necessary to conduct research and monitor the use of this drug in plasma and cerebrospinal fluid (CSF) patients with TBM. This study aimed to determine the method of analysis of INH and PZA using Ultra Performance Liquid Chromatography Ultra Violet (UPLC-UV). The samples were taken from TBM patients who taken INH-PZA and others besides TBM patients who did not take INH-PZA. The analytic method carried out includes a comparison of the results of the analysis method for measuring levels of standard INH-PZA and INH-PZA in plasma and CSF samples. Analysis of INH-PZA in plasma and CSF can be performed using UPLC with UV detector, at least 100 uL plasma or CSF samples volume, with reproducible and accurate results.

A Novel Ultra Performance Liquid Chromatographic Method for the Estimation of Lercanidipine in Bulk and Tablet Dosage Form

Aim: In this present study an accurate reverse phase ultra-performance liquid chromatography (RP-UPLC) method has been developed, validated and applied to stability indicating studies to determine Lercanidipine HCL in bulk and marketed dosage form.&#x0D; Methods: Optimized chromatographic conditions were achieved by using Waters Acquity BEH C18 (2.1 x 50mm, 1.7m) UPLC column. Empower 2 is a software, dihydrogen Orthophosphate Buffer : Methanol (40 : 60) as eluent at flow rate 0.3 ml/min. PDA detection was performed at 254nm.&#x0D; Results: The developed method was validated and stability study was conducted as per ICH guidelines. The retention time was found at 0.503 min. The method shows linearity over a range of 1 μg/ml to 60 μg /ml with the obtained correlation coefficient is 0.999. The LOD and LOQ values were found 0.025 and 0.05 μg /ml. The acidic and peroxide stressed study shows more degradation of 6.23% and 3.03%.&#x0D; Conclusion: The present developed method was found stability indicating, reliable, validated method was applied for the routine analysis of lercanidipine in bulk drug and the pharmaceutical formulations.

ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF COMBINATION OF ANTI-ASTHMATIC DRUGS MONTELUKAST AND DOXOFYLLINE

For qualitative and quantitative analysis, various analytical techniques are available such as Ultraviolet (UV) Spectrophotometry, High-performance liquid chromatography (HPLC), High-performance thin layer chromatography (HPTLC). As per literature survey, there are some UV, HPLC, Ultra-Performance Liquid Chromatography (UPLC) and HPTLC analytical methods are developed for Montelukast and Doxofylline individually and in a combination with other drugs too, since yet there are no significant stability studies indicating HPLC method reported for Montelukast and Doxofylline combinations. In the current study, the HPLC method is developed and validated for simultaneous quantitative estimations of Montelukast and Doxofylline. These present techniques are more efficient and sensitive as compared to other analytical techniques.